Days Of Incubation Research Articles

Developmental toxicity of short-chain chlorinated paraffins on early-stage chicken embryos in a shell-less (ex-ovo) incubation system

Short-chain chlorinated paraffins (SCCPs) are listed as a category of globally controlled persistent organic pollutants (POPs) by the Stockholm Convention in 2017. However, SCCP toxicity, particularly their developmental toxicity in avian embryos, has not been well studied. In this study, we observed the early development of chicken embryos (Gallus gallus domesticus) by applying a shell-less (ex-ovo) incubation system developed in our previous studies. After exposing embryos at Hamburger Hamilton stage (HHS) 1 to SCCPs (control, 0.1% DMSO; SCCPs-L, 200 ng/g; SCCPs-M, 2000 ng/g; SCCPs-H, 20,000 ng/g), we observed the development of embryos from the 3rd to 9th incubation day. Exposure to SCCPs-M and -H induced a significant reduction in survival, with an LD50 of 3100 ng/g on the 9th incubation day. Significant dose-dependent decreases in body length were observed from days 4–9. We also found that SCCPs-H decreased the blood vessel length and branch number on the 4th incubation day. Additionally, SCCPs-H significantly reduced the heart rate on the 4th and 5th incubation days. These findings suggest that SCCPs may have potential of developmental and cardiovascular toxicity during the early stages of chicken embryos. Quantitative PCR of the mRNA of genes related to embryonic development showed that SLC16A10 (a triiodothyronine transporter) level decreased in the SCCPs-H group, showing a significant positive correlation with the body length of embryos. THRA level, a thyroid hormone receptor, was significantly decreased in the SCCPs-H group, whereas that of DIO3 level, a deiodinase was significantly increased. These results suggest that SCCPs exposure induces developmental delays via the thyroxine signaling pathway. Analysis of thyroid hormones (THs) in blood plasma also indicated a significant reduction in thyroxine (T4) levels in the SCCPs-H group on the 9th incubation day of embryos. In conclusion, SCCPs induce developmental toxicity by disrupting thyroid functions at the early-life stage of chicken embryos.

Microbial response to long-term fertilization of paddy soils: Apparent and real priming effects

Fertilization is crucial for increasing crop productivity and it alters soil microbial biomass and activities. These alterations exert implications for soil carbon (C) stocks, primarily through the priming effect (PE). Here, we investigated the mechanisms underlying PE and their impact on soil C stocks in paddy soils subjected to long-term (31 years) fertilization. Soils from four depths within the upper 40 cm layer, subjected to four fertilization types (no fertilizer, mineral fertilizer, mineral + straw, mineral + chicken manure), were incubated for 60 days with or without 13C-labeled glucose. Following substrate addition, an increase in isotopically unlabeled CO2 efflux may stem from accelerated microbial turnover (apparent PE) or modified SOM mineralization (real PE). By analyzing microbial biomass (MBC), CO2 and dissolved organic C (DOC) dynamics, we explored the partitioning of C from glucose into three pools. Following glucose addition (day 2), the soil organic matter (SOM)-derived MBC decreased by 4.7 % at 0–10 cm in unfertilized and by 6.3 % in mineral-fertilized soils. SOM-derived MBC also decreased in soils fertilized with straw- and chicken manure at 0–20 and 0–30 cm. Hence, the apparent PE closely correlated with fertilization types. At day 2, the SOM-derived DOC increased with the increase in extractable Fe2+ content, indicating high contribution of the released mineral-bound organic C in the DOC pool. Microbial decomposition of SOM-derived DOC and loss of unlabeled C through apparent PE increased the release of unlabeled CO2, resulting in a positive but low PE. After 60 days of incubation, the addition of glucose to the subsoils (30–40 cm) caused a net loss of C (positive PE) of 6.6–19 mg kg−1. In contrast, negative PEs were observed in the topsoil, indicating a decrease in microbial activity and C turnover. These findings suggested that fertilization regimes, whether involving long-term extra C input or not, can alter microbial biomass and turnover rates. Moreover, the C turnover mechanisms in paddy soils are complex, including apparent PE, release of mineral-bound organic C, and negative PE. Collectively, these processes can consequently change soil C stocks.

Development of Plumeria alba extract supplemented biodegradable films containing chitosan and cellulose derived from bagasse and corn cob waste for antimicrobial food packaging

With the increase in global plastic pollution due to conventional plastic packaging (petroleum-derived), bioplastics have emerged as an alternative green source for practising a circular economy. This research aimed to extract cellulose from bagasse and corn cob waste and utilized in mixed form to prepare bioplastic film. The mixed cellulose was further reinforced with natural substances such as chitosan, bentonite, and P. alba extract. These newly developed bioplastics films were characterized by various physical tests like film thickness, moisture content, water solubility and spectroscopic techniques such as Fourier transform infrared (FTIR), scanning electron microscopy-energy dispersive spectroscopic (SEM-EDX), thermal gravimetric analysis (TGA), and ultraviolet-visible (UV–Vis) spectroscopy for opacity testing. The results revealed the enhanced bioplastic thermal and mechanical characteristics through robust interactions between cellulose and bentonite molecules. Moreover, incorporating chitosan solution as reinforcements in bio-composite films resulted in improved water barrier properties. The results indicated lower absorption in the UV range of 250–400 nm, attributed to the absence of UV-absorbing groups. Finally, their biodegradability was tested in soil, and 85.3 % weight loss of bioplastic films was observed after 50 days of the experiment which is the main task of this research. The antimicrobial properties of bioplastic films have been evaluated, and showed an inhibition zone of 16 mm against E. coli. After 12 days of incubation of sherbet berries, complete spoilage is identified in the control group compared to those covered with the bioplastic film. This outcome is attributed to the antioxidant and antimicrobial activities provided by chitosan and P. alba extract in the bioplastic film. The comprehensive outcomes of this study suggest the potential future adoption of these entirely bio-derived, environmentally sustainable and biodegradable bioplastic films as a viable substitute for the plastic packaging currently present in the market.

Virulence Genes and Biofilm Formation Among Legionella pneumophila Isolates Collected from Hospital Water Sources.

Legionella pneumophila can be transmitted to people, especially immunocompromised patients, via hospital water pipe systems and cause severe pneumonia. The aim of our study was to investigate the presence of major virulence factor genes, ability of biofilms formation, and correlation between presence of Legionella isolates and temperature, pH, and residual chlorine of water. Hundred water samples were collected from nine hospitals in Tehran, Iran. Temperature, pH, and residual chlorine were determined during sampling. Different virulence genes and the ability to form biofilms were subsequently analyzed among the L. pneumophila isolates. Results showed that 12 (12%) samples were positive in culture method and all of the isolates were positive as L. pneumophila species (mip). A correlation was found between Legionella culture positivity and temperature and pH of water, but there was no significant correlation between residual chlorine of water samples and the presence of Legionella. The isolation of Legionella rate in summer and spring was higher than winter and autumn. Twelve (100%) isolates were positive for mip genes, 9 (75%) for dot genes, 8 (66.66%) for hsp, 6 (50%) for lvh, and 4 (33.33%) for rtx. All of the isolates displayed strong ability for biofilm production every three days. Two of these isolates (16.6%) displayed weak ability to form biofilm on the first day of incubation. This study revealed that water sources in hospitals were colonized by virulent Legionella and should be continuously monitored to avoid elevated concentrations of Legionella with visible biofilm formation.

Embryotoxicity of fluconazole on developing chick embryos

Abstract Background Fluconazole is a first-generation triazole used as an antifungal treatment for skin, hair, and nail infections. The study aimed to assess the embryotoxicity and teratological effects of fluconazole on chick embryos. Fertilized eggs were divided into four groups: two experimental groups treated with different concentrations of fluconazole (0.1 ml/egg, 0.2 ml/egg) containing 12 and 11 eggs, respectively, one group treated with distilled water (10 eggs), and a control group (10 eggs) left untreated. The liquid form of fluconazole was administered on the 4th day of incubation, and observations were made on the 9th day. Results There were different anomalies observed in them like hydrocephaly, microcephaly shorting of the beak, agenesis, Amelia, micromelia, anophthalmia, microphthalmia, and kyphosis. There were also observed the morphometric measurements with a difference of significant (p < 0.001) and (p < 0.01) in CR length, body weight, head circumference, eye circumference, forelimb, and hind limb. Different vital organ defects were also observed in histological studies. Fluconazole in various combinations has been found to cause embryotoxicity and teratological consequences in chick embryos. Conclusion The current study showed that fluconazole is teratogenic in creating chick undeveloped organisms. The embryotoxic impacts were recorded on structural morphometric and graphic levels.

Effects of Land-Use Type and Salinity on Soil Carbon Mineralization in Coastal Areas of Northern Jiangsu Province

Sea level rise due to glacier melting caused by climate warming is a major global challenge, but the mechanism of the effect of salinity on soil carbon (C) mineralization in different land types is not clear. The pathways by which salinity indirectly affects soil carbon mineralization rates need to be investigated. Whether or not the response mode is consistent among different land-use types, as well as the intrinsic links and interactions between soil microbial resource limitation, environmental stress, microbial extracellular enzyme activity, and soil carbon mineralization, remain to be demonstrated. In this paper, three typical land-use types (wetland, forest, and agroforestry) were selected, and different salinity levels (0‰, 3‰, 6‰, and 32‰) were designed to conduct a 125-day laboratory incubation experiment to determine the soil CO2 release rate, soil physicochemical properties, and soil enzyme activities, and to correlate C mineralization with biotic and abiotic factors. A correlation analysis of soil physical and chemical properties, extracellular enzyme activities, and carbon mineralization rates was conducted to investigate their intrinsic linkages, and a multiple linear regression of C mineralization at different sites was performed to explore the variability of mineralization among different site types. Structural equation models were established in the pre- and post-incubation stages to study the pathways of soil C mineralization at different incubation times, and the mechanism of mineralization was further verified by enzyme stoichiometry. The results showed that, at the end of 125 days of incubation, the 32‰ salinity addition reduced the cumulative mineralization of forest and agroforestry types by 28.41% and 34.35%, respectively, compared to the 0‰ salinity addition. Soil C mineralization in the three different land-use types was highly correlated with the active C fractions of readily oxidizable C (ROC), dissolved organic C, and microbial biomass C (MBC) in the soil, with the standardized coefficients of multivariate linear regression reaching 0.67 for MBC in the wetland and −0.843 for ROC in the forest. Under long-term salinity additions, increased salinity would reduce the microbial respiratory quotient value by inhibiting β-glucosidase activity, thus indirectly affecting the rate of CO2 release. With added salinity, the mineralization of non-saline soil was more susceptible to the inhibitory effect of salinity, whereas the mineralization of salinized soil was more controlled by soil C pools.

Degradation of textile polyazodyes by Polyporus ciliatus laccase produced in peanut shell solid medium: purification and characterization

This study describes decolorization of reactive sulfonated azoic dye Direct Blue 71, one of the recalcitrant and highly used industrial dyes, by Polyporus ciliatus derived laccase. Agro-industrial wastes are increasingly used as substrates for laccase production from white rot fungi through solid-state fermentation (SSF). Peanut shell (PS) is an attractive substrate for laccase production, and SSF is an adequate strategy for that purpose. It has been employed as a novel support-substrate for laccase production by Polyporus ciliatus under solid-state fermentation. The highest laccase yield of 1260 U/ml was attained using PS after 13 days of incubation at 30°C. The purified laccase (55 kDa) showed maximum activity at pH 3 and 60°C. The extracellular laccase enzyme was purified 67.6-fold to a specific activity of 1466 U/mg by a Mono-Q anion-exchange column. The purified enzyme Km value was found to be 24 µM, while the Vmax value was observed to be 0.96 µM s-1 with ABTS as a substrate. It had high thermotolerance and pH stability and was resistant to several metal ions including copper, cadmium, and iron. P. ciliatus pure laccase showed high potential for the decolorization and detoxification of the reactive sulfonated azoic dye Direct Blue 71, which suggested that this enzyme could be used for textile effluent treatment.

Outbreak of Septoria leaf spot caused by Sphaerulina musiva on Populus × euramericana in China.

In June 2023, severe leaf spots were noted in Populus × euramericana cv 'Nanlin95' plantations located in the Nanjing Baguazhou Wetland Park (32°09'16.97″N, 118°48'16.74″E) of Jiangsu Province and Populus × canadensis cv 'Sacrau 79' and Populus × canadensis cv 'Guariento' in the Liyuan Village in Nanyang City (32°53'43.70″N, 112°17'29.12″E) of Henan Province, respectively. The disease incidence in both locations could reach 97.9% (556 out of 568 trees) and 98.9% (2409 out of 2436 trees), respectively. The initial symptoms appear as numerous small and circular spots (1.59 to 3.18 mm in diameter) with gray or tan centers and dark-brown margins on the leaves. As the spots age, they sometimes enlarge, often coalesce, and may extend down the petioles. Diseased leaves and petioles were both surface sterilized with 75% ethanol for 30 seconds. With the aid of a hand lens, pycnidia (brown to black, spherical in profile, 90 to 250 µm diam) were easily picked out in the center of the spots and subsequently transferred into 1 mL sterilized water for preparing the spore suspension plated on KV8 medium amended with 100 mg/liter streptomycin sulfate and 50 mg/liter chloramphenicol. After 12 days of incubation, 86 single-spore isolates were obtained and identified as typical Septoria-like fungi according to morphological features, including slow-growing, gray or black colonies with pink mucilaginous matrix and hyaline, straight or curved conidia (size = 25 to 59 × 3.5 to 4 μm; septa = 1 to 6). Species identification was further validated by PCR amplification and sequencing of the internal transcribed spacer (ITS) region with ITS1/ITS4 primer pairs. Multiple sequence alignments with ClustalW revealed that the obtained ITS sequences of 86 isolates were 100% identical to each other. A BLAST search in GenBank indicated that the selfsame sequences of two representative isolates (isolate BGZ11 of Jiangsu Province, accession no. OR660379; isolate KZB22 of Henan Province, accession no. OR711499) shared 99.8% identity (494 of 495 bp) and 100% identity (504 of 504 bp) with related sequences of Sphaerulina musiva (Peck) Quaedvlieg, Verkley, and Crous (syn. = Septoria musiva Peck) in GenBank (MN275187; KF251619), respectively. Furthermore, we used a S. musiva-specific PCR assay (Abraham et al. 2018) on symptomatic leaf samples collected from the plantation. Each sample consisted of 20 cut-out leaf spots per leaf. Eight of the 10 samples were positive for S. musiva DNA. To confirm pathogenicity, six sterile tissue culture of poplar plants (Populus trichocarpa and Populus × euramericana cv 'Nanlin895') were respectively transplanted into pots and grown in a greenhouse for a week and for a month with an 18-h photoperiod augmented with sodium lamps and a 20°C (day)/16°C (night) temperature regime. Inoculations were conducted by spraying the plants with conidia suspension (106 conidia/mL) (LeBoldus et al. 2010). Control plants were sprayed with distilled water. Leaf spots were developed on the inoculated P. trichocarpa leaves at one week and P. × euramericana cv 'Nanlin895' leaves at 10 days after inoculation while no symptoms were observed on the control plants. The fungus S. musiva was successfully reisolated from all symptomatic leaves fulfilling Koch's postulates. Sphaerulina musiva only causes an endemic leaf spot disease on its natural North American host Populus. deltoides (Feau et al. 2010; Ostry 1987). However, on susceptible Populus species (e.g., P. balsamifera, P. trichocarpa, P. maximowiczii) and hybrids, S. musiva causes not only leaf spots but also severely damaging stem and branch cankers (Jeger et al. 2018; LeBoldus et al. 2009; Sondreli et al. 2020). To our knowledge, this is the first report of S. musiva causing leaf spots on poplar in China. Large-scale timber imports (e.g., cut branches, isolated bark, wood with and without bark) potentially lead to anthropogenic-facilitated transport of this pathogen. This outbreak of Septoria leaf spot underscores the potential threat of this pathogen to P. × euramericana in China, where it is widely planted as a keystone forestry species.

Allantoic Fluid-Based qPCR for Early Onset In Ovo Sexing.

The day-old male chick culling remains a welfare issue in the poultry industry. Several governments have prohibited this practice, pushing hatcheries to seek alternatives. Although different solutions exist for solving this problem, sex determination during the embryo's incubation (in ovo sexing) is considered the most suitable one among the consumers and industry. However, to be industrialized, in ovo sexing technologies must meet several requirements: compatibility with all egg colors and early developmental stages while maintaining a high hatchability rate and accuracy at low cost and high throughput. To meet these requirements, we studied the use of the sexual genes HINTW (female-specific) and DMRT-1 (both sexes) at incubation days 6-9. By utilizing the quantitative polymerase chain reaction in allantoic fluid (AF) samples, our study confirmed female-specific HINTW detection on all days without any significant detrimental effects on embryo development. We achieved 95% sexing accuracy using the HINTW cycle threshold (Ct) alone and 100% accuracy rate when using Δλ values (difference between the HINTW and DMRT-1 Ct). In conclusion, the developed assay can provide information about AF as a sample for in ovo sexing and open new industrial possibilities for faster and cheaper assays.

Comparison of Human Long-Term Liver Models for Clearance Prediction of Slowly Metabolized Compounds.

The accurate prediction of human clearance is an important task during drug development. The proportion of low clearance compounds has increased in drug development pipelines across the industry since such compounds may be dosed in lower amounts and at lower frequency. Such compounds present new challenges to in vitro systems used for clearance extrapolation. In this study we compared the accuracy of clearance predictions of suspension culture to four different long-term stable in vitro liver models, including HepaRG sandwich culture, the Hµrel stochastic co-culture, the Hepatopac micropatterned co-culture (MPCC) and a micro-array spheroid culture. Hepatocytes in long-term stable systems remained viable and active over several days of incubation. Although intrinsic clearance values were generally high in suspension culture, clearance of low turnover compounds could frequently not be determined using this method. Metabolic activity and intrinsic clearance values from HepaRG cultures were low and, consequently, many compounds with low turnover did not show significant decline despite long incubation times. Similarly, stochastic co-cultures occasionally failed to show significant turnover for multiple low and medium turnover compounds. Among the different methods, MPCCs and spheroids provided the most consistent measurements. Notably, all culture methods resulted in underprediction of clearance, this could however be compensated for by regression correction. Combined, the results indicate that spheroid culture as well as the MPCC system provide adequate in vitro tools for human extrapolation for compounds with low metabolic turnover. Significance Statement In this study, we compared suspension cultures, HepaRG sandwich cultures, the Hµrel liver stochastic co-cultures, the Hepatopac micropatterned co-cultures (MPCC) and micro-array spheroid cultures for low clearance determination and prediction. Overall, HepaRG and suspension cultures showed modest value for the low determination and prediction of clearance compounds. The micro-array spheroid culture resulted in the most robust clearance measurements, whereas using the MPCC resulted in the most accurate prediction for low clearance compounds.

Internal Transcribed Spacers (ITS) Based Identification of Trichoderma Isolates and Biocontrol Activity against Fusarium oxysporum and Chitinase Activity

Twelve Trichoderma spp. isolated from different locations in Uttar Pradesh, India. Internal transcribed spacer (ITS) amplification and sequencing were used to identify isolates of Trichoderma that showed 99–100% identification with Trichoderma harzianum, Trichoderma asperellum, and Trichoderma longibrachiatum, the three species of Trichoderma. Tests were conducted in Vitro to evaluate the biocontrol potential of Trichoderma isolates against Fusarium species. After seven days of incubation, the isolate TBT6 (T. harzianum) exhibited the greatest antagonism against Fusarium oxysporum, with inhibition rates of 87.1%) by TBT7 (82.2%), and the least amount of inhibition (59.7%) by T. longibrachiatum isolate TBT10. Under field conditions, the antagonistic T. harzianum isolate TBT6 can be employed as a bio-control agent against Fusarium oxysporum. and for the development of formulations based on Trichoderma. Using the dual plate approach, twelve highly antagonistic Trichoderma isolates were selected for chitinolytic activity. Observing the breakdown of chitin substrates, Trichoderma isolates TBT6 was shown to have strong chitinolytic enzyme activity, making it a good candidate for endochitinase gene isolation. Using two particular primers, the genomic DNA of the Trichoderma isolate TBT6 was isolated and amplified.

Controlling organic carbon increase in oxygenated marine sediment by using decarburization slag

The chemical oxygen demand (COD) in the Seto Inland Sea, Japan has increased in the recent decades due to the increase of bottom dissolved oxygen (DO) concentration which stimulated several autotrophic microorganisms, specially sulfur oxidizing bacteria (SOB). This increased SOB activity due to the oxygenation of the bottom sediment synthesized new organic matter (OM) which contributed dissolved organic carbon to the overlying seawater. This phenomenon further led to hypoxia in some subareas in the Seto Inland Sea. Higher pH or alkaline environment has been found to be an unfavorable condition for SOB. In this research, we used decarburization slag to elevate the pH of sediment to control the SOB activity and consequently reduce OM production in the sediment. Ignition loss of the surface sediment increased from 5.14% 6.38% after 21 days of incubation with aeration; whereas the sediment showed the less ignition loss of 5.71% after 21 days when the slag was incubated in the same experimental setup. Microbial community analysis showed less SOB activity in the slag added aerated sediment which accounts for the controlled increase of OM in the sediment. An additional experiment was conducted with magnesium oxide to confirm whether elevated pH can control the OM increase in sediment due to rising DO. All these results showed that decarburization slag can elevate the pH of the sediment to a certain level which can control the SOB activity followed by controlled increase of OM in the sediment. The findings may be beneficial to control accumulation of sedimentary OM which can act as a source of organic carbon in the overlying seawater.

Influence of combined abiotic/biotic factors on decay of P. aeruginosa and E. coli in Rhine River water

Understanding the dynamic change in abundance of both fecal and opportunistic waterborne pathogens in urban surface water under different abiotic and biotic factors helps the prediction of microbiological water quality and protection of public health during recreational activities, such as swimming. However, a comprehensive understanding of the interaction among various factors on pathogen behavior in surface water is missing. In this study, the effect of salinity, light, and temperature and the presence of indigenous microbiota, on the decay/persistence of Escherichia coli and Pseudomonas aeruginosa in Rhine River water were tested during 7 days of incubation with varying salinity (0.4, 5.4, 9.4, and 15.4 ppt), with light under a light/dark regime (light/dark) and without light (dark), temperature (3, 12, and 20 °C), and presence/absence of indigenous microbiota. The results demonstrated that light, indigenous microbiota, and temperature significantly impacted the decay of E. coli. Moreover, a significant (p<0.01) four-factor interactive impact of these four environmental conditions on E. coli decay was observed. However, for P. aeruginosa, temperature and indigenous microbiota were two determinate factors on the decay or growth. A significant three-factor interactive impact between indigenous microbiota, temperature, and salinity (p<0.01); indigenous microbiota, light, and temperature (p<0.01); and light, temperature, and salinity (p<0.05) on the decay of P. aeruginosa was found. Due to these interactive effects, caution should be taken when predicting decay/persistence of E. coli and P. aeruginosa in surface water based on a single environmental condition. In addition, the different response of E. coli and P. aeruginosa to the environmental conditions highlights that E. coli monitoring alone underestimates health risks of surface water by non-fecal opportunistic pathogens, such as P. aeruginosa.Key pointsAbiotic and biotic factors interactively affect decay of E. coli and P. aeruginosaE.coli and P.aeruginosa behave significantly different under the given conditionsOnly E. coli as an indicator underestimates the microbiological water quality

Effects of in ovo injection of organic selenium on the hatchability of broiler breeder hen eggs and resulting chick physiology and performance.

Selenium is an essential mineral for poultry. The conflicting reports about its in ovo injection are the justification for the more detailed investigation. The aim of this study was to investigate the effects of in ovo injection of organic selenium on the hatching traits of broiler chickens and their performance. Three hundred and twenty eggs of Ross 308 strain with an average weight of 65g and 160 chicks were randomly divided into 4 treatment groups (each with 8 replicates of 10 eggs each for hatching parameters and 4 replicates of 10 chicks for broiler farming parameters): negative control (no injection), positive control (in ovo injection of 0.272mL of normal saline solution) and 2 selenium treatments (in ovo injection of 2.72 or 5.44μg of organic selenium). Injection was into the amniotic sac on the 10th day of incubation. Effects of in ovo injection on hatching and performance traits, blood parameters, immune responses, carcass characteristics, meat fatty acid profile, cecal microbial population and selenium consternation in the tibia were measured. Fewer chicks from the injected treatments hatched than from the negative control group (p<0.01). However, the injection of selenium increased feed intake and the final weight of the birds (p<0.01). Blood parameters were also affected. Glucose and cholesterol in experimental treatment chicks was lower than those of the controls (p<0.01), whereas blood lipoproteins (VLDL, LDL and HDL) and the ratio of cholesterol to HDL was significantly increased in the treatments injected with selenium (p<0.01). There was no significant difference in the immune response or microbial population between the experimental groups, but carcass components, such as thigh, breast, wing and abdominal fat weight, were significantly greater in the selenium treatments. Intra-egg injection of organic selenium produced favourable effects on performance of broiler chickens, although it had no effect on immune response or microbial population. However, the negative effect on hatching of chickens needs to be prevented to result in an acceptable economic return for the producer.

Adenosine triphosphate induces amorphous aggregation of amyloid β by increasing Aβ dynamics

Amyloid β (Aβ) aggregates into two distinct fibril and amorphous forms in the brains of patients with Alzheimer’s disease. Adenosine triphosphate (ATP) is a biological hydrotrope that causes Aβ to form amorphous aggregates and inhibit fibril formation at physiological concentrations. Based on diffracted X-ray blinking (DXB) analysis, the dynamics of Aβ significantly increased immediately after ATP was added compared to those in the absence and presence of ADP and AMP, and the effect diminished after 30 min as the aggregates formed. In the presence of ATP, the β-sheet content of Aβ gradually increased from the beginning, and in the absence of ATP, the content increased rapidly after 180 min incubation, as revealed by a time-dependent thioflavin T fluorescence assay. Images of an atomic force microscope revealed that ATP induces the formation of amorphous aggregates with an average diameter of less than 100 nm, preventing fibrillar formation during 4 days of incubation at 37 °C. ATP may induce amorphous aggregation by increasing the dynamics of Aβ, and as a result, the other aggregation pathway is omitted. Our results also suggest that DXB analysis is a useful method to evaluate the inhibitory effect of fibrillar formation.

Morphogenetic Promoters in the Development of Provisional and Definitive Urinary Organs in Humans and Vertebrates

The aim is to show the regulatory significance of promoter organs of morphogenesis in a stable state of the kidneys of vertebrates (fish, birds) and humans. Material and methods. 118 embryos at stages 12-23 of Carnegie (SC) and 28 human fetuses of 9-12 weeks of fetogenesis, 268 embryos of meat chickens (cross "Hibro PG +") from the stage of 48 hours to 20 days of broiler egg incubation, 50 fry of peled Coregonus peled were studied (Gmelin, 1788) at the stage of yolk sac atrophy (stage 36-37 according to the classification of Detlaf, 1975). To study the method of light microscopy, the material fixed in a 10% neutral form, embedded in paraffin. Sections stained with Mayer's hematoxylin and eosin, McManus CHIC method. Ki-67, CD31, CD34, positive cells are detected by immunohistochemistry. Primary kidney examination by low-vacuum electron microscopy. Embryos and human fetuses are observed in the Tyumen health care facility during medical abortion on demand in anamnestic healthy women with their warning about emergency work. Observation chicken embryos at the poultry farm “Tyumensky Broiler”, incubation workshop (Kaskara village, Tyumen region). Morphometric analysis and statistical processing of the actual material were carried out. Results. It was shown that the organotypic differentiation of the intermediate mesenchyme and the formation of nephrons of the provisional and definitive urinary organs control the state of the mesonephric duct and metanephritic diverticulum. The mesonephric duct initiates the organotypic antibiotic of the perifocal intermediate mesonephric mesenchyme and detects the proximodistal vector of saltatory mesonephronogenesis. The metanephritic diverticulum and its daughter branches initiate the organotypic differentiation of the intermediate mesenchyme and control the fan mechanism of metanephronogenesis. Activity of nephronogenesis Determination of the dynamics of convergence of immunocompetent cells. The absence of ventrodorsal generation of mesonephrons in humans is associated with a significant vital cycle of a normal kidney, atrophy of the mesonephric duct, and a decrease in the activity of immunocompetent cells. An increase in the expression of CD31, CD34, Bcl-2, Ki-67 positive cells is realized in combination with the formation of nephron rudiments as the promoter organs move in the developing organ of urinary formation. Conclusion. Prenatal organogenesis monitors and constantly monitors the formation of promoter organs of morphogenesis - the mesonephric duct and the metanephric disserticulum. Organotypic infection of the intermediate mesenchyme during the development of fish, birds and humans correlates with the chronovector of the convergence of immunocompetent cells.

Ecological Significance of Dynamism of Physiologic Variance on Properties of Palm Oil Mill Effluents

Properties a given waste is essential for rationale comprehension of plausible way(s) for its remediation. In the present study, POME from oil milling centre at Anambra state showed high heterotrophic activity of microorganisms. Total viable counts of the organisms isolated from the waste water were 5.4 x 103 CFU/ml and 4.4 x 103 CFU/ml at day 0 and day 7 of the incubation respectively. These observations showed a significant variation from the control experiment at these recorded days of incubations which shows total viable cell counts (TVC) of 4.6 x 103 and 2.9 x 104 CFU/ml respectively. Total coliform counts (TCC) of the organisms isolated from the dairy waste water were 1.3 x 103 and 1.8 x 103 CFU/ml for the day 0 and day 7 incubation periods respectively while the TCC from the control experiment were: 8.0 x 102 and 3.2 x 102 CFU/ml for day 0 and day 7 incubation periods respectively. POME sample from Awka, Anambra state was subjected to various physicochemical profiling and was further optimized at different conditions to exert various variability. Biochemical oxygen demand (BOD5) is one of the important physical parameter widely used in assessment of quality of aqueous surroundings. Physicochemical properties of waste water from the milling centre showed the following: pH (5.0), temperature (41oC), conductivity (610); water dissolved minerals such as Cl-, K+, PO3, Mg2+, Ca2+ were all determined. Initial dissolved oxygen concentration (mg/l) of the waste water was determined using a probe connected to a meter with initial value of 39.4 mg/l. Biochemical oxygen demand (BOD5) was determined after five incubation days at room temperature in separate bottles optimized at varying pH of 4.0-9.0 in a range of 1.0 units. BOD5 decreases with increased pH values in all the tested samples incubated for 5 days. pH 5.0 gave a significant reduction of the oxygen exertion (1400 mg/ml) while pH 4.0, 6.0, 7.0 showed an oxygen exertion (BOD5) of 1978, 2778, and 2965 mg/ml respectively. Samples incubated at 8.0 and 9.0 showed oxygen exertion of 2945 and 2899 mg/ml respectively. Other contents of the dairy waste water such as total organic carbon (TOC), total organic matter contents (TOM) were all determined respectively. Samples was also collected from mapped area 0.5 km away from the waste water reservoir and used as the reference standard. Proceeding results from the present research study showed the compromised statues of our aquatic environment and also in a wider look gave a great insight of effective modalities in water treatment; as increasing population of myriads of processing industries in our country today pose great challenge to the competence of our aquatic body.

Exploration and Optimization of Potential Fungi to Degrade Herbicides with Active Ingredient Isopropylamine Glyphosate from Shallot Plantations by In Vitro

This research was conducted by isolating potential fungi from soil samples exposed the herbicides with the active ingredient of isopropylamine glyphosate (IAG) and then testing the degradation ability of the obtained fungal isolates. The results showed 15 isolates fungi obtained from the soil to a depth of 10 cm including Aspergillus sp. with black, green and yellow colonies, Cladosporium sp., Penicillum sp., Rhizopus sp., Mucor sp., Cephalosporium sp., Phymatotrichum sp., Pytopthora sp., Curvularia sp., Microsporum sp., Colletothricum sp., Acremonium sp., Tricophyton sp. and 4 isolates from soil samples with a depth of 20 cm, namely Chrysosporium sp., Geotrichum sp., Aspergillus sp. with black and green colonies. However, after testing for the presence of herbicides with concentrations of 10 ppm, 20 ppm, and 30 ppm, only 2 isolates were able to survive, namely Aspergillus sp with black and green colonies. Further testing on the ability to degrade the IAG was found to reduce the concentration of herbicide. The optimum biodegradability to IAG of Aspergillus sp. green and black found in P20 (20 ppm) treatment at the 5th day of incubation, namely 98.69 ppm for Aspergillus sp. green, and 96.11 ppm for Aspergillus sp. black. Keywords: Aspergillus sp., Biodegradation, Fungi, Herbicide, Isoprofile amine glyphosate.

Noninvasive Assessment of Chicken Egg Fertility during Incubation Using HSSE-GC-MS VOC Profiling.

Volatile organic compounds (VOCs) carry crucial information about chicken egg fertility. Assessing the fertility before incubation holds immense potential for poultry industry efficiency. Our study used headspace sorptive extraction-gas chromatography-mass spectrometry to analyze egg VOCs before and during the initial 12 incubation days. A total of 162 VOCs were identified. Hexanal was significantly higher in unfertilized eggs, whereas compounds such as propan-2-ol, propan-2-one, and carboxylic acids were higher in fertilized eggs. Furthermore, the obtained multiple logistic regression model outperformed the partial least-squares-discriminant analysis (PLS-DA) model, demonstrating lower complexity and superior performance. Fertile eggs were accurately identified in the validation set in 68-75% of the cases during the initial 4 days, to 85 and 100% on days 6 and 8. Finally, hierarchical cluster analysis in fertilized eggs revealed the clustering of VOCs of the same chemical class, indicative of their shared biochemical origin. This suggests a promising direction for future research aimed at understanding the biological information embedded in VOCs and their relationship to biochemical processes during embryo development.

First Report of Black Leaf Spot Caused by Epicoccum latusicollum on Chinese Yam in China.

Chinese yam (Dioscorea polystachya Turczaninow cv. Tiegun), which belongs to the family Dioscoreaceae, is widely cultivated throughout China due to its high economic and medicinal value. In June 2023, black leaf spots on Chinese yam (cv. Purple 1) were observed in Nanchang city (28.45° N, 115.49° E) of Jiangxi province, southeastern China. The incidence of the disease ranged between 70 and 85% of plants, and up to 30% of the leaves per plant were affected in the field over a 2-week period of study. Infected foliage displayed brown necrotic lesions, elliptical or irregular, with yellow halo at the edge of the lesion (0.5 to 3 cm diam.). To identify the causal agent, 32 symptomatic leaves of eight symptomatic plants were collected. Small pieces from the margin of necrotic leaf tissue (about 3 x 3 mm) were surface sterilized in 75% ethanol for 30 s followed in 0.1% HgCl2 for 1 min, and washed three times with ddH2O. Then, the pieces were transferred onto potato dextrose agar (PDA) plates and incubated at 26°C for 3 days with a 12-h light-dark cycle. From the 32 isolates, 21 exhibited similar morphology after hyphal tipping resulting in an isolation frequency of 65.6%. Colonies on PDA were initially white aerial hyphae but became grayish with age, and a reddish orange pigment on the underside. After 16 days of incubation, pycnidia were observed, which were dark, spherical or flat spherical, and 64.1 to 172.5 µm (n = 25) in diameter. Conidia were ellipsoidal, aseptate, hyaline, and 4.1 to 5.6 × 1.8 to 2.7 µm (n = 80). In addition, a blackish green discoloration was produced on malt extract agar (MEA) using the NaOH spot test. The isolates were tentatively identified as Epicoccum spp. based on morphological characteristics (Chen et al. 2017). Isolate AYZ-1 was randomly selected for identification and pathogenicity testing. Genomic DNA of the isolate (AYZ-1) was extracted and amplified by polymerase chain reaction (PCR) using ITS1/ITS4 for the internal transcribed spacer (ITS) region (White et al. 1990), Btub2Fd/Btub4Rd for the β-tubulin (TUB) region (Woudenberg et al. 2009), LROR/LR7 for the large ribosomal RNA gene (LSU) region (Rehner and Samuels 1994), and RPB2-5F2/fRPB2-7cR for RNA polymerase II second largest subunit (RPB2) region(Liu et al. 1999), respectively. The concatenated sequences (GenBank Accession No. OR574165, OR567827, OR574166, OR567828, respectively) shared 99.8 to 100% identity withEpicoccum latusicollum(OP788080, MN329871, OR428532, and OL422485, respectively). A neighbor-joining phylogenetic tree was generated based on the concatenated sequences in MEGA7, placed isolate (AYZ-1) within E. latusicollum. To fulfill Koch's postulates, healthy leaflets from three one-year-old Chinese yam (cv. Purple 1) were used as inoculation materials, using isolate AYZ-1. Two sites of each leaf were wounded with a sterile needle and covered with a piece of cotton drenched with 200 µL spore suspension (106 conidia/mL) on the left sides, while sterilized water served as the control on the right sides of leaves. All inoculated leaves were covered with clear polyethylene bags for 24 h. Plants were grown outdoors at a daily average temperature of 26°C with relative humidity over 45%. After 7 days of incubation, the leaves showed the same symptoms as the original diseased leaves. The E.latusicollumisolate was re-isolated from diseased leaves and confirmed by morphology and sequencing analysis, fulfilling Koch's postulates. E.latusicollumhas beenpreviously reportedtocauseblack root on yam inChina'ssouth-westernprovince ofSichuan(Han et al. 2019). Meanwhile,leafspothave been reportedon many plants by this genus, such astobacco(Guo et al. 2020) andbanana(Liu et al. 2023). According to our knowledge, this is thefirstreportofE.latusicollumcausingblack leafspoton Chinese yam inChina. This finding will provide an important reference for understanding the biology of E. latusicollum and the distribution of the disease, but more research is needed to determine if management is warranted.